3D cell culture model for compound screening

We have developed a novel 3-D cell culture system for cell based screening and phenotypic drug discovery in a high throughput multi-well plate setting.

Unlike other 3-D cell biology systems, this new technology (patent pending) was developed to offer the unique ability to accommodate any cell type in any assay at any throughput. The same system can be used to develop and validate assays and allows transition to high throughput screening capacities.

Traditional 2D cell culture has been the mainstay for basic biology and drug discovery for decades. The development of this micro-scaffold based 3D cell culture system recapitulates the structural basis of the extracellular matrix and provides a platform for cells to adhere and proliferate on, representing a more physiologically relevant micro-environment. This novel 3D microscaffold technology has proven to be compatible with all common biological plate based assay types including; luminescence, fluorescence, high content imaging and real-time kinetic studies.

Our 3D technology, based on electrospun material (Electrospinning Company, Harwell, Oxford, UK) in a micro-scaffold architecture paves the way to a paradigm shift in screening from simple 2-D to more amenable 3-D based higher throughput assays and, if coupled with iPSC’s and differentiated stem cells or human derived primary cells further enhances a phenotypic approach to drug discovery, introducing a more physiological environment to cell-based HTS system.

Figure 1: Scanning electron micrograph of a single 3D micro-scaffold

Figure 2:  Lightsheet confocal microscopy image of a number of 3D micro-scaffolds seeded with epithelial cells

Key points;

  • Robust and reproducible 3D culture environment
  • Applicable to any cell type (recombinant, primary and iPSC)
  • Pipettable and scalable to any assay throughput
  • Integrates seamlessly with all current screening and assay workflows

Figure 3: Recombinant HEK-293 cells expressing a cyclic AMP (cAMP) response element (CRE) fused to firefly luciferase were cultured for 48 hours on micro-scaffolds then treated in well plates with forskolin. Forskolin produces a dose-dependent increase in cAMP within cells detected as an increase in luminescence signal. The response measured from cells cultured in 2-D matches that of cells cultured in 3-D and suggests that similar pharmacology can be obtained between 2-D and 3-D cells.

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